Mesenchymal stem cells and their regenerative and immunomodulatory potential Abstract Mesenchymal stem cells (MSCs) possess multidirectional regenerative ability, which, together with their immunomodulatory potential, makes them promising cell type for therapy of wide variety of diseases. Despite ongoing research, which proved MSCs application to be safe, reported effect of MSCs administration on patients is not convincingly beneficial yet. In our work we focused on elucidation of MSCs role in regeneration of vital organs, heart and liver, where a large damage is life threatening for patients and any improvement in therapy would save many lives. Similar situation is in Graft versus host disease (GVHD), where MSCs immunomodulatory properties could be beneficial. Role of MSCs in heart regeneration was examined in vitro. Primary adult swine cardiomyocytes (CMCs) were co-cultured with or without swine MSCs for 3 days and morphological and functional parameters (contractions, current, respiration) of CMCs were measured. MSCs showed supportive effect on CMCs survival, especially at day 3 of the experiment, where in co-culture was significantly higher number of viable CMCs with physiological morphology and maintained function. Effect of MSCs on liver regeneration was observed in swine model of chronic liver...

1 Abstract Liver fibrosis is a condition described by extensive accumulation of scar tissue in the liver. With further progression, it leads to cirrhosis or even to hepatocellular carcinoma. Liver fibrosis accompanies every chronic liver disease and its prevalence in adult European population is estimated to be around 4%. During my dissertation work, I studied the function of three members of Metzincin family of metalloproteinases - ADAM17, ADAM10 and MMP-19, in liver fibrosis and liver regeneration using mouse genetic models. ADAM17 and ADAM10 are important regulators of signalling pathways which are involved in immune response as well as differentiation. Both proteases are able to cleave ectodomains of their substrates from cell membrane, affecting bioavailability of ligands and functionality of receptors. Several of their substrates are involved in liver pathologies. MMP-19 on the other hand, is a metalloprotease mainly involved in extracellular matrix cleavage, important process in fibrosis development, as well as resolution of fibrosis. Our results demonstrate that ablation of ADAM10 results in increased susceptibility to liver fibrosis in mice, both spontaneous and toxin induced. ADAM10 deficiency affected biliary epithelium, as we detected higher markers of biliary damage in serum of ADAM10 deficient...

Iron is an essential element for human organism, because it cooperates as a cofactor of enzymes in many metabolic pathways. Iron is a component of hemoglobin, and thus it is indispensable for the oxygen transport to tissues. It can exist as a ferrous or ferric form. However, ferrous iron paticipates in reactions in which highly reactive hydroxyl group can be formed. This product is harmful for the organism. Non-heme iron is taken up to the circulation through duodenal enterocyte. Iron excretion is carried out only by desquamation of the enterocytes or by bleeding. Therefore, iron intake must be strictly regulated. Iron overloading is observed in some chronic diseases (hereditary hemochromatosis, alcohol liver disease). In contrary, iron depletion can be a case of iron deficiency anemia. The aim of this master thesis is to determine the expression of iron transport molecules in duodenum in chronic diseases which originate due to disturbances of iron intake regulation. We determine the expression of molecules of iron transport (DMT1, Dcytb, ferroportin, hephaestin) on mRNA level by qPCR and on protein level by western blot. The level of serum hepcidin was determined by ELISA. Our results show an increased expression of mRNA of transporters DMT1 and ferroportin as well as ferrireductase Dcytb and ferroxidase...

This diploma thesis deals with home parenteral nutrition in the day and night regime and its effect on patients. Parenteral nutrition is usually administered at night so that patients can do normal daily activities and so that nutrition administered in this way has the least impact on their lives. Parenteral nutrition is not a physiological route of nutrient administration and it is not natural for the human body to process nutrients at night. The first part of the theoretical work presents parenteral nutrition and its composition, indications, contraindications and complications. The next part of the thesis describes home parenteral nutrition and its organisation which is necessary for this form of nutritional support to be implemented. It is necessary to monitor many laboratory values on parenteral nutrition, the theoretical part specifically mentions cholesterol, triacylglycerols, glycemia or liver function tests. One of the important parts is the chapter about circadian rhythms, hormones and metabolic changes in energy intake at night. Qualitative research was chosen for this work, which involved 17 patients on home parenteral nutrition. These patients were getting their nutrition during the night. Their laboratory values such as liver function tests, glycaemia and blood fats was evaluated and...

Mesenchymal stem cells and their regenerative and immunomodulatory potential Abstract Mesenchymal stem cells (MSCs) possess multidirectional regenerative ability, which, together with their immunomodulatory potential, makes them promising cell type for therapy of wide variety of diseases. Despite ongoing research, which proved MSCs application to be safe, reported effect of MSCs administration on patients is not convincingly beneficial yet. In our work we focused on elucidation of MSCs role in regeneration of vital organs, heart and liver, where a large damage is life threatening for patients and any improvement in therapy would save many lives. Similar situation is in Graft versus host disease (GVHD), where MSCs immunomodulatory properties could be beneficial. Role of MSCs in heart regeneration was examined in vitro. Primary adult swine cardiomyocytes (CMCs) were co-cultured with or without swine MSCs for 3 days and morphological and functional parameters (contractions, current, respiration) of CMCs were measured. MSCs showed supportive effect on CMCs survival, especially at day 3 of the experiment, where in co-culture was significantly higher number of viable CMCs with physiological morphology and maintained function. Effect of MSCs on liver regeneration was observed in swine model of chronic liver...

Lowering of central venous pressure in hepatic surgery is nowadays widely recommended and used procedure. Low central venous pressure anesthesia is associated with decreased blood loss and improved clinical outcome. There are several approaches how to reach low central venous pressure. Till now none of them is recommended as superior in terms of patient safety and clinical outcome. Concurrently there is still debate if to use the low central venous pressure anesthesia principle or if it could be replaced with a principle of anesthesia with high stroke volume variation (or another dynamic preload parameter) with the use of a more sophisticated hemodynamic monitoring method. Results of our study didn't show any significant difference between two approaches used for reduction of central venous pressure, but suggest that the principle of low central venous pressure anesthesia could be possibly replaced by the principle of high stroke volume variation anesthesia which presumes the use of advanced hemodynamic monitoring. KEYWORDS Hepatic resection, central venous pressure, Pringle maneuver, hemodynamics, hemodynamic monitoring, fluid therapy, anesthesia

A stationary or portable infusion pump can be used to administer home parenteral nutrition (HPN). The stationary pump (SP) must be fasten to the infusion stand, so patients administer infusions in night mode to maintain their quality of life and mobility in the daytime. However, administering the HPN at nighttime can cause complications. Conversely, the portable pump (PP) does not have to be fasten to the infusion stand and the infusions can be administered in day mode during activities, as the PP can be placed in a special backpack. The aim of this diploma thesis is to evaluate the administration of HPN in day mode in patients with a PP and further to evaluate and compare the results of quality of life and liver function tests results in patients with a PP and a SP. HPN patients with a PP and a SP completed the HPN-QOL questionnaire in the nutritional clinic at the 4th internal clinic of the General University Hospital in Prague during outpatient visits. In addition to that, patients with a PP completed a questionnaire focused on the administration of HPN in day mode. Liver function tests results (total bilirubin, ALT, AST, GGT, ALP, cholinesterase and albumin) were obtained from patients' medical records. A total of 14 respondents with a PP and 14 respondents with a SP were included in the...

This thesis focuses on the importance of heme catabolic pathway in hepatic carcinogenesis. Hepatocellular carcinoma (HCC) is the most common primary malignant liver tumour. It is primarily caused by hepatic cirrhosis or chronic viral hepatitis. Hepatic carcinogenesis is associated with increased oxidative stress. Thus, our study aimed to assess the expression of the genes involved in the homeostasis of oxidative stress in patients with HCC. The study was performed on patients with primary HCC (n = 29) and control subjects (n = 11), gene expressions of heme oxygenase 1 (HMOX1), biliverdin reductase A/B (BLVRA/B), endothelial growth factor (VEGFA), NADPH oxidase 2 (NOX2) and p22phox were analyzed in the liver tissue; the identical gene expressions were analyzed in peripheral blood leukocytes (PBL) in HCC patients (n = 32) and in healthy controls (n = 27). Expression of BLVRA in liver tissue and PBL was significantly increased in cancer patients, as was NOX2 expression in PBL. Expression of VEGFA in liver tissue and PBL was significantly decreased in cancer patients. The expression of the other genes studied did not differ between HCC patients and controls. These data are consistent with the results of our previous study on patients with chronic hepatitis C, in which BLVRA expression was also...

Lowering of central venous pressure in hepatic surgery is nowadays widely recommended and used procedure. Low central venous pressure anesthesia is associated with decreased blood loss and improved clinical outcome. There are several approaches how to reach low central venous pressure. Till now none of them is recommended as superior in terms of patient safety and clinical outcome. Concurrently there is still debate if to use the low central venous pressure anesthesia principle or if it could be replaced with a principle of anesthesia with high stroke volume variation (or another dynamic preload parameter) with the use of a more sophisticated hemodynamic monitoring method. Results of our study didn't show any significant difference between two approaches used for reduction of central venous pressure, but suggest that the principle of low central venous pressure anesthesia could be possibly replaced by the principle of high stroke volume variation anesthesia which presumes the use of advanced hemodynamic monitoring. KEYWORDS Hepatic resection, central venous pressure, Pringle maneuver, hemodynamics, hemodynamic monitoring, fluid therapy, anesthesia

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Kryštof Dobečka Supervisor: PharmDr. Tomáš Smutný, Ph.D. Advisor: prof. PharmDr. Petr Pávek, Ph.D. Title of diploma thesis: Experimental approaches for studying hepatic enzyme induction mediated by pregnane X receptor The thesis focuses on hepatic pregnane X receptor (PXR)-mediated induction of biotransformation enzymes. Emphasis is placed on experimental models and methods which are used for the assessment of enzyme induction. In addition to summarizing its well- established role as a xenobiotic-sensing receptor, PXR is also presented as a transcription factor with an important role in endogenous pathways. Furthermore, cell and animal models are evaluated in terms of expression and function of PXR and its target xenobiotic-metabolising enzymes. Primary human hepatocytes in 2D cultures are considered to be the gold standard of in vitro hepatic models. However, 3D technologies are expected to be increasingly used in the future. The use of animal models is limited due to pronounced interspecies differences in PXR activation. Thus, humanized models have been established to overcome these limitations. Next, this thesis comments screening methods for an assessment of interaction between PXR and...